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The purpose of this study was purification and characterization of phenol monooxygenase from Stenotrophomonas maltophilia strain KB2, enzyme that catabolises phenol and its derivatives through the initial hydroxylation to catechols. The enzyme requires NADH and FAD as a cofactors for activity, catalyses hydroxylation of a wide range of monocyclic phenols, aromatic acids and dihydroxylated derivatives of benzene except for catechol. High activity of this monooxygenase was observed in cell extract of strain KB2 grown on phenol, 2-methylphenol, 3-metylphenol or 4-methylphenol. Ionic surfactants as well as cytochrome P450 inhibitors or 1,4-dioxane, acetone and n-butyl acetate inhibited the enzyme activity, while non-ionic surfactants, chloroethane, ethylbenzene, ethyl acetate, cyclohexane, and benzene enhanced it. These results indicate that the phenol monooxygenase from Stenotrophomonas maltophilia strain KB2 holds great potential for bioremediation.
Nowadays biodegradations of harmful xenobiotics seems to be the best and cheapest method of purification of the polluted environment. VOCs are represented by vinyl acetate, which is thought to be carcinogenic. The aim of these studies was to isolate and determine the susceptibility profile for vinyl acetate of bacterial strains. The source of microorganisms was soil sampled in the area of Synthos S.A. in Oświęcim, Poland. From among 41 isolates, 4 Gram-negative strains were chosen for further analyses. As the control, one laboratory strain of Pseudomonas fluorescens PCM 2123 from The Polish Collection of Microorganisms (Wrocław) was used. Simultaneously, a susceptibility profile to vinyl acetate was performed on Stenotrophomonas maltophilia KB2 strain, aromatic compounds’ degrader. Vinyl acetate used in concentration of 3,000 ppm inhibited growth of gram-positive bacteria, and 4,000 ppm was the lethal dose for microorganisms from mixed populations. A toxicity test showed susceptibility to vinyl acetate at concentrations of 2,000 ppm. Three weeks of pre-incubation with 400 ppm of vinyl acetate magnified the level of sensitivity to 3,000 ppm of vinyl acetate for almost all strains. Although decomposition of vinyl acetate was observed even in the presence of 4,000, 5,000 and 6,000 ppm of vinyl acetate, growth was not observed. It was due to enlarged concentration of acetaldehyde, a product of hydrolysis ester bond of vinyl acetate.
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